OPTICAL AND PHOTOPHYSICAL PROPERTIES OF THE OXAZOLE YELLOW DNA PROBES YO AND YOYO

Artikel i vetenskaplig tidskrift, 1994

The photophysical properties of the optical DNA probe YOYO (homodimeric derivative of oxazole yellow) have been characterized in terms of the monomeric part, the YO chromophore. In aqueous solutions YO is virtually nonfluorescent but upon binding to DNA its fluorescence quantum yield is strongly increased. A similar enhancement of the fluorescence is observed for YO in the viscous solvent glycerol. The high fluorescence quantum yield of YO, when bound to DNA or in a viscous solution, is proposed to be a result of decreased rotational mobility around the internuclear bridge between the two aromatic ring systems. This hypothesis is based on similar values of the activation energies for the temperature-dependent nonradiative decay processes (E(A) = 53 kJ/mol) and viscous flow (E(A) = 63 kJ/mol), suggesting related rate-limiting mechanisms. A single electronic transition is found to be responsible for the intense visible absorption band. This conclusion is based on the observation of an essentially wavelength-independent reduced linear dichroism and similarly wavelength independent fluorescence anisotropy, and the fact that the emission spectrum is very nearly a mirror image of the absorption spectrum. The conclusion is further supported by quantum mechanical calculations (CNDO/S). By combination of measurements of fluorescence anisotropy of YO in glycerol and linear dichroism of YO in a stretched poly(vinyl alcohol) film, the transition moment of the strong visible absorption band was found to be nearly long axis polarized, in agreement with the CNDO/S calculations. The low-energy electronic transition and its polarization direction in the YO chromophore remain essentially unperturbed in the YOYO dye, suggesting that the results obtained for the excited state of the YO chromophore are applicable also to YOYO. One difference, though, is that in aqueous solutions the two YO chromophores of YOYO interact with each other, forming an internal dimer, resulting in a distorted absorption spectrum.